This invention relates to a series of small molecules which bind to the erythropoietin receptor and compete with the natural ligand for binding to said receptor. The invention includes pharmaceutical compositions containing these mimetics, their methods of production as well as intermediates used in their synthesis.
Erythropoietin (EPO) is a 34,000 dalton glycoprotein hormone which is produced in the mammalian kidney. Its primary role is stimulation of mitotic cell division and differentiation of erythrocyte precursor cells. As a result this hormone regulates the production of erythrocytes, the hemoglobin contained therein and the blood""s ability to carry oxygen. The commercial product Epogen(copyright) is used in the treatment of anemia. This drug is produced by recombinant techniques and is formulated in aqueous isotonic sodium chloride/sodium citrate. Even though it has been used successfully in the treatment of anemia, it is a costly drug that is administered intravenously. This method of administration is both costly and inconvenient for the patient; therefore it would be desirable to find a EPO mimetic which has the potential for oral activity.
A small molecule EPO mimetic has advantages over the natural protein. The immune response associated with large peptides is unlikely to occur with small molecules. In addition, the variety of pharmaceutical formulations that may be used with small molecules are technically unfeasible for proteins. Thus the use of relatively inert formulations for small molecules is possible. The most important advantage of small molecules is their potential for oral activity. Such an agent would ease administration, cost less and facilitate patient compliance.
Although compounds which mimic EPO are useful in stimulating red blood cell synthesis, there are diseases where the overproduction of red blood cells is a problem. Erythroleukemia and polysythemia vera are examples of such diseases. Since EPO is an agent responsible for the maturation of red blood cell precursors, an antagonist of EPO would have utility treating either of those diseases.
The disclosed invention consists of a series of small molecules which demonstrate competitive binding with the natural ligand for the EPO receptor. As such these compounds are potentially useful in the treatment of diseases or conditions associated with this receptor. In addition, the invention contemplates methods of producing these compounds and intermediates used in their production.
The invention includes compounds of the Formula I: 
wherein:
R1 is the side chain of a natural or unnatural ax-amino acids, where if said side chain contains a protectable group, that group may be protected with a member of the group consisting of succinyl, glutaryl, 3,3-dimethylglutaryl, C1-5alkyl, C1-5alkoxycarbonyl, acetyl, N-(9-fluorenylmethoxycarbonyl), trifluoroacetyl, omega-carboxyC1-5alkylcarbonyl, t-butoxycarbonyl, benzyl, benzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, phenylsulfonyl, ureido, t-butyl, cinnamoyl, trityl, 4-methyltrityl, 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, tosyl, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, phenylureido, and substituted phenylureido (where the phenyl substituents are phenoxy, halo, C1-5alkoxycarbonyl);
R2 and R3 
may be taken together to form a six-membered aromatic ring which is fused to the depicted ring, or
are independently selected from the group consisting of hydrogen, C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, amino, phenyl, phenoxy, phenylC1-5alkyl, phenyl C1-5alkoxy,
substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted phenoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted phenylC1-5alkyl (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted phenylC1-5alkoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), and
substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halo substituted C1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substitutents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
R4and R5 
may be taken together to form a six-membered aromatic ring which is fused to the depicted ring, or
are independently selected from the group consisting of hydrogen, C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, amino, phenyl, phenoxy, phenylC1-5alkyl, phenyl C1-5alkoxy,
substituted phenyl (where the substituents are selected from C1-5alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted phenoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted phenylC1-5alkyl (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino),
substituted phenylC1-5alkoxy (where the substituents are selected from C1-5 alkyl, C1-5 alkoxy, hydroxy, halo, trifluoromethyl, nitro, cyano, and amino), and
substituted amino (where the substituents are selected from one or more members of the group consisting of C1-5alkyl, halosubstitutedC1-5alkyl, C1-5alknyl, C1-5alkenyl, phenyl, phenylC1-5alkyl, C1-5alkylcarbonyl, halo substituted C1-5alkylcarbonyl, carboxyC1-5alkyl, C1-5alkoxyC1-5alkyl, cinnamoyl, naphthylcarbonyl, furylcarbonyl, pyridylcarbonyl, C1-5alkylsulfonyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, phenylC1-5alkylsulfonyl substituted phenylcarbonyl, substituted phenylC1-5alkylcarbonyl, substituted phenylsulfonyl, substituted phenylC1-5alkylsulfonyl, substituted phenyl, and substituted phenylC1-5alkyl [where the aromatic phenyl, phenylC1-5alkyl, phenylcarbonyl, phenylC1-5alkylcarbonyl, phenylsulfonyl, and phenylC1-5alkylsulfonyl substitutents are independently selected from one to five members of the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino]);
W is selected from the group consisting of xe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94Sxe2x80x94, and xe2x80x94CHxe2x95x90Nxe2x80x94;
Q is selected from the group consisting of xe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94Sxe2x80x94, and xe2x80x94CHxe2x95x90Nxe2x80x94;
X is selected from the group consisting of carbonyl, C1-5alkyl, C1-5alkenyl, C1-5alkenylcarbonyl, and (CH2)mxe2x80x94C(O)xe2x80x94 where m is 2-5;
Y is selected from the group consisting of carbonyl, C1-5alkyl, C1-5alkenyl, C1-5alkenylcarbonyl, and (CH2)mxe2x80x94C(O)xe2x80x94 where m is 2-5;
n is 1, 2, or 3;
Z is selected from the group consisting of hydroxy, C1-5 alkoxy, phenoxy, phenylC1-5alkoxy, amino, C1-5alkylamino, diC1-5alkylamino, phenylamino, phenylC1-5alkylamino, piperidin-1-yl
substituted piperidin-1-yl (where the substituents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, halo, aminocarbonyl, C1-5alkoxycarbonyl, and oxo;
substituted phenylC1-5alkylamino (where the aromatic substitutents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, phenylC1-5alkenyloxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino),
substituted phenoxy (where the aromatic substitutents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino),
substituted phenylC1-5alkoxy (where the aromatic substitutents are selected from the group consisting of C1-5alkyl, C1-5alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, and amino), xe2x80x94OCH2CH2(OCH22CH2)sOCH2CH2Oxe2x80x94, xe2x80x94NHCH2CH2(OCH2CH2)sOCH2CH2NHxe2x80x94, xe2x80x94NH(CH2)pO(CH2)qO(CH2)pNHxe2x80x94, xe2x80x94NH(CH2)qNCH3(CH2)sNHxe2x80x94, xe2x80x94NH(CH2)sNHxe2x80x94, and (NH(CH2)s)3N, where s, p, and q are independently selected from 1-7
with the proviso that if n is 2, Z is not hydroxy, C1-5 alkoxy, amino, C1-5alkylamino, diC1-5alkylamino, phenylamino, phenylC1-5alkylamino, or piperidin-1-yl, with the further proviso that if n is 3, Z is (NH(CH2)s)3N.
and the salts thereof.
The terms used in describing the invention are commonly used and known to those skilled in the art. xe2x80x9cIndependentlyxe2x80x9d means that when there are more than one substituent, the substitutents may be different. The term xe2x80x9calkylxe2x80x9d refers to straight, cyclic and branched-chain alkyl groups and xe2x80x9calkoxyxe2x80x9d refers O-alkyl where alkyl is as defined supra. xe2x80x9cCbzxe2x80x9d refers to benzyloxycarbonyl. xe2x80x9cBocxe2x80x9d refers to 1-butoxycarbonyl and xe2x80x9cTsxe2x80x9d refers to toluenesulfonyl. xe2x80x9cDCCxe2x80x9d refers to 1,3-dicyclohexylcarbodiimide, xe2x80x9cDMAPxe2x80x9d refers to 4-Nxe2x80x2,N-dimethylaminopyridine and xe2x80x9cHOBTxe2x80x9d refers to 1-hydroxybenzotriazole hydrate. xe2x80x9cFmocxe2x80x9d refers to N-(9-fluorenylmethoxycarbonyl), xe2x80x9cDABCOxe2x80x9d refers to 1,4-Diazabicyclo[2.2.2]octane, xe2x80x9cEDCIxe2x80x9d refers to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, xe2x80x9cDdexe2x80x9d refers to 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl, and xe2x80x9cTMOFxe2x80x9d refers to trimethyl orthoformate. The side chains of xcex1-amino acids refer to the substituents of the stereogenic carbon of an xcex1-amino acid. For example if the amino acid is lysine, the side chain is 1-aminobutan-4-yl. The term natural amino acid refers to the 20 xcex1-amino acids of the L configuration which are found in natural proteins. Unnatural xcex1-amino acids include synthetic amino acids such as, xcex1-aminoadipic acid, 4-aminobutanoic acid, 6-aminohexanoic acid, xcex1-aminosuberic acid, 5-aminopentanoic acid, p-aminophenylalanine, xcex1-aminopimelic acid xcex3-carboxyglutamic acid, p-carboxyphenylalanine, carnitine, citrulline, xcex1,xcex2-diaminopropionic acid, xcex1,xcex3-diaminobutyric acid, homocitrulline, homoserine, and statine as well as D-configuration amino acids. The term xe2x80x9cprotectable groupxe2x80x9d refers to a hydroxy, amino, carboxy, carboxamide, guanidine, amidine or a thiol groups on an amino acid side. Compounds of the invention may be prepared by following general procedures known to those skilled in the art, and those set forth herein.
The compounds of the invention may be prepared by liquid phase organic synthesis techniques or by using amino acids which are bound to a number of known resins. The underlying chemistry, namely, acylation and alkylation reactions, peptide protection and deprotection reactions as well as peptide coupling reactions use similar conditions and reagents. The main distinction between the two methods is in the starting materials. While the starting materials for the liquid phase syntheses are the N-protected amino acids or the lower alkyl ester derivatives of either the N-protected or N-unprotected amino acids, the starting material for the resin syntheses are N-protected amino acids which are bound to resins by their carboxy termini.
An equivalent of an N-Fmoc-protected amino acid which is bound to a resin 1a is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. A solution of about two to three equivalents of an appropriately substituted halide, 1b, and a suitable base such DIEA is added to the resin bound amino acid and this mixture is shaken for 18-36 h. The resulting mixture is washed with several portions of a suitable solvent and is suspended and shaken in an acidic solution, such as 50% TFA/CH2CH2, over several hours to cleave the acid from the resin and give the N-disubstituted amino acid 1c.
By varying the resin bound amino acid 1a, one may obtain many of the compounds of the invention. The following resin bound amino acids may be used in Scheme I: alanine, N-g-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)arginine, xcex2-(4-methyltrityl)asparagine, aspartic acid (xcex2-t-butyl ester), S-(trityl)cysteine, xcex3-(4-methyltrityl)glutamine, glutamic acid (xcex2-t-butyl ester), glycine, N-imidazolyl-(trityl)histidine, isoleucine, leucine, N-xcex5-(2-chlorobenzyloxycarbonyl)lysine, N-xcex5-(t-butoxycarbonyl)lysine, methionine, phenylalanine, proline, O-(t-butyl)serine, O-(t-butyl)threonine, N-indolyl-(t-butoxycarbonyl)tryptophan, O-(t-butyl)tyrosine, valine, xcex2-alanine, xcex1-aminoadipic acid, 4-aminobutanoic acid, 6-aminohexanoic acid, xcex1-aminosuberic acid, 5-aminopentanoic acid, p-aminophenylalanine, xcex1-aminopimelic acid xcex3-carboxyglutamic acid, p-carboxyphenylalanine, carnitine, citrulline, xcex1,xcex2-diaminopropionic acid, xcex1,xcex3-diaminobutyric acid, homocitrulline, homoserine, and statine. In addition, the choice of xe2x80x9cWxe2x80x9d and xe2x80x9cXxe2x80x9d can be varied by using known halide derivatives of 1b. For example using benzylchloride, 2-chloromethylthiophene, or 2-chloromethylpyridine gives compounds of the invention where xe2x80x9cWxe2x80x9d is xe2x80x94CHxe2x95x90CHxe2x80x94, xe2x80x94Sxe2x80x94, or xe2x80x94CHxe2x95x90Nxe2x80x94, respectively. For variations in xe2x80x9cXxe2x80x9d, the use of 2-chloroethylphenyl, 3-chloro-1-propenylbenzene, or benzeneacetyl chloride as 1b, give compounds where Y is (CH2)2, xe2x80x94CHxe2x95x90CHxe2x80x94CH2xe2x80x94, or xe2x80x94CH2C(O)xe2x80x94 respectively. Still further, Scheme 1 may be used to produce combinatorial mixtures of products. Using mixtures of resin bound amino acids, 1a, with only one 1b produces said combinatorial mixtures. Alternatively, using one amino acid 1a with a mixture of 1b as well as mixture of 1a with mixtures of 1b gives a large range of combinatorial mixtures. 
An equivalent of an N-Fmoc-protected amino acid which is bound to a resin 1a is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. Trimethyl orthoformate and an appropriately substituted aldehyde 2a (5 equivalents) is added and the mixture is shaken under N2 overnight. This mixture is treated with a suspension of NaBH(OAc)3 (5 equivalents) in CH2Cl2 and shaken under N2 overnight. After filtration and washing with a suitable solvent, the resulting product, resin bound Nxcex1-monosubstituted amino acid 2b, is rinsed with a suitable solvent and its identity is confirmed by MS and or HPLC analysis after treatmet of a portion of the resin with 50% TFA/CH2Cl2.
The resin 2b is suspended in an appropriate solvent such as DMF and is filtered. The appropriately substituted alkyl or arylkyl halide, 2c, and an appropriate base such as DIEA are added with some additional solvent and the mixture is shaken under N2 for 18-36 h. The resin bound Nxcex1,Nxcex1-disubstituted amino acid, 2d, is isolated from the suspension and the resin is cleaved with an acidic solution to give the free acid 2e. 
A resin bound amine, 2d, where R4 is nitro, is suspended in a suitable solvent, such as DMF, and is filtered. This mixture is treated with SnCl2 dihydrate in DMF and shaken under N2 overnight. The solvent is removed and the resin is washed successive portions of a suitable solvent to give the resin bound compound 3a where R4 is amino. The resin is suspended in a suitable solvent and is combined with an organic base, such as pyridine an appropriately substituted carboxylic acid anhydride, acid chloride, or sulfonyl chloride. The mixture is shaken under N2 overnight and is filtered to give the resin bound amino acid 3b. This material is treated with an acid and a suitable solvent to give the free amino acid 3b.
The resin bound amine 3a is treated with TMOF and an appropriately substituted aldehyde 3c is added and the mixture is shaken under N2 overnight. The resulting mixture is drained and treated with a suspension of NaBH(OAc)3 in an appropriate solvent and this mixture is shaken under N2 overnight. The resin bound 3-aralkylaminophenyl amino acid is identified my spectral techniques after clevage to give the free acid 3d as previously described.
Resin bound, 2d, where R1 is (CH2)4NH(Dde) is mixed with a suitable solvent, such as DMF, and shaken with successive portions of 2% solution of hydrazine hydrate in DMF over about 30 min. The resin is filtered and treated with a suitable solvent and a cyclic anhydride derivative 3e, and a base such as DMAP and pyridine. This mixture is shaken under N2 overnight and filtered to give the resin bound amine, 3f. This material is identified by spectral techniques after clevage to give the free acid 3f as previously described. 
Resin bound 2b, where R2 is nitro is suspended in CH2Cl2 and is treated with an organic base, such as pyridine, and 9-fluorenylmethoxy chloride. This mixture is shaken under N2 overnight, filtered and resuspended in a suitable solvent. This mixture is treated with SnCl2 dihydrate in DMF and shaken under N2 overnight. The solvent is removed and the resin is washed successive portions of a suitable solvent and filtered to give the resin bound compound 4a where R2 is amino. The resin 4a is then suspended in a suitable solvent, such as CH2Cl2, and is combined with 0.4 mmol of pyridine and 0.25-0.4 mmol of the appropriately substituted carboxylic acid anhydride, acid chloride, or sulfonyl chloride. The mixture is shaken under N2 overnight, filtered, and washed successively with three portions each of CH2Cl2 and MeOH. This resin is suspended in DMF, filtered, and shaken under N2 with 5 mL of a 40% solution of piperidine in DMF. After 1 h, the solvent is drained and the resin was washed successively with three portions each of suitable solvents to give the resin bound 4b. The identity of the compound was confirmed by spectral analysis after cleveage as previously described. 
The resin 2b (0.2 mmol) is suspended in CH2Cl2, filtered, and is resuspended in CH2Cl2. This suspension is treated with diethyl phosphonoacetic acid and diisopropylcarbodiimide or other suitable carbodiimide reagent, and the mixture is shaken under N2 overnight. The solvent is drained and the resulting resin 5a was washed successively with three portions each of CH2Cl2 and MeOH. The resin is suspended in DMF and filtered. A solution of the appropriately substituted aldehyde 5b (0.6-1.0 mmol) in 3-5 mL of DMF, lithium bromide (0.6-1.0 mmol), and a suitable base such as DIEA or Et3N (0.6-1.0 mmol) is added and the mixture is shaken under N2 overnight. The solvent is removed and the resin is washed successively with three portions each of DMF, CH2Cl2, and MeOH. The identity of the resin bound substituted amino acid 5c was confirmed spectral techniques. The resin bound material may be treated with 50% TFA/CH2Cl2 over 1-1.5 h, to give the acid 5c. 
To prepare compounds where n is 2 and Z is NH(CH2)sNH, products of Schemes 1-5 may be used in Scheme 6. Treatment of two equivalents of the substituted amino acid 1c with an equivalent of the diamine 6a, in the presence of HOBT and a peptide coupling agent such as EDCI and a base such as DIEA at room temperature over 16 h gives the dimer 6b. 
A solution of of amino acid ester 7a, an appropriately substituted halide derivitive 1b, and an appropriate base such as DIEA, Na2CO3, or Cs2CO3 in a suitable solvent, such as DMF, is heated at 50-100xc2x0 C. under N2 overnight, or until the starting material is exhausted, to give a mixture of the di and mono-substituted amines, 7b and 7c respectively. If the side chains of R1 contain acid cleavable protecting groups, those groups may be cleaved by treatment with 30-80% TFA/CH2Cl2. Esters 7b and 7c may be independently converted to the corresponding acids 7d and 7e by hydrolysis with an appropriate base such as aqueous NaOH. 
A solution of 1 mmol of amino acid ester 8a (or the corresponding HCl salt and 1.1 mmol of DIEA) and 1-1.5 mmol of the appropriately substituted aldehyde 2a in 3-5 mL of trimethyl orthoformate was stirred at room temperature under N2 overnight. The solution was either concentrated and used directly for the next reaction, or was partitioned between EtOAc and water, washed with brine, dried over Na2SO4, and concentrated to give crude product, which was purified by MPLC to give mono-substituted product 8b.
Amino ester 8b was dissolved in DMF, combined with 1.1-1.5 mmol of the appropriately substituted chloride or bromide 2c, and heated at 50-100xc2x0 C. overnight. The reaction mixture was cooled and partitioned between water and EtOAc. The organic layer was washed three times with water and once with brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure 8c. For examples of 8c wherein the side chain R1 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 8c was stirred in 30-80% TFA/CH2Cl2 for 1-3 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 8c. For examples of 8c where R9 was equal to t-butyl, 8c was stirred in 30-80% TFA/CH2Cl2 for 1-3 h and treated as described above to give acid 8d. For examples of 8c where R9 was equal to methyl, ethyl, or other primary or secondary alkyl esters, 8c was stirred with with 1-2 mmol of aqueous LiOH, NaOH, or KOH in MeOH, EtOH, or THF at 20-80xc2x0 C. until TLC indicated the absence of 8c. The solution was acidified to pH 4-5 with aqueous citric acid or HCl and was extracted with CH2Cl2 or EtOAc. The organic solution was washed with brine, dried over Na2SO4, and concentrated to give 8d.
For examples of amino acid ester 8c where R1=(CH2)4NHBoc, 8c (1 mmol) was stirred in 30-80% TFA/CH2Cl2 for 1-3 h. The reaction mixture was concentrated to provide 8e as the TFA salt. Optionally, the TFA salt was dissolved in CH2Cl2 or EtOAc and washed with aqueous NaOH or Na2CO3, dried over Na2SO4, and concentrated to give 8e as the free base.
A solution of 1 mmol of 8e, 1-4 mmol of an appropriate base such as DIEA, and 1-2 mmol of the appropriately substituted cyclic anhydride 3e was stirred in CH2Cl2 or DMF under N2 overnight. The resulting mixture was diluted with CH2Cl2 or EtOAc and washed with aqueous HCl, water, and brine, was dried over Na2SO4, and concentrated to provide 8f. Alternatively, 1 mmol of 8e, 1-4 mmol of an appropriate base such as DIEA, and 1-2 mmol of the appropriately substituted carboxylic acid anhydride (R11CO)2O or acid chloride R11COCl was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 8g. Alternatively, 1 mmol of 8e, 1-4 mmol of an appropriate base such as DIEA, and 1-2 mmol of the appropriately substituted isocyanate R12NCO was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 8h. 
For examples of 8c where R5=NO2, a solution of 1 mmol of 8c (where R2, R3, R4, or) and 10-12 mmol of SnCl2 dihydrate was stirred in MeOH, EtOH, or DMF at 20-80xc2x0 C. for 0.5-24 h under N2. The solution was taken to room temperature and poured into aqueous Na2CO3 with rapid stirring. The resulting mixture was extracted with EtOAc or CH2Cl2 and the organic extracts were washed with brine, dried over Na2SO4, and concentrated to give the aminophenyl product 9a, which was purified by MPLC or used without further purification.
A solution of 1 mmol of aminophenyl compound 9a and 1-1.5 mmol of the appropriately substituted aldehyde 2a in 3-5 mL of trimethyl orthoformate was stirred at room temperature under N2 overnight. The solution was either concentrated and used directly for the next reaction, or was partitioned between EtOAc and water, washed with brine, dried over Na2SO4, and concentrated to give crude product, which was purified by MPLC to give 9b. For examples of 9b wherein the side chain R1 or R9 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 9b was stirred in 30-80% TFA/CH2Cl2 for 1-3 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 9b.
A solution of 1 mmol of 3-aminophenyl compound 9a, 1.1-2 mmol of pyridine, and 1-1.5 mmol of the appropriately substituted acid chloride, acid anhydride, or sulfonyl chloride in 3-5 mL of CH2Cl2 or ClCH2CH2Cl was stirred at room temperature under N2 overnight. The solution was partitioned between EtOAc and water, washed with water, saturated aqueous NaHCO3, and brine, dried over Na2SO4, and concentrated to give crude product which was optionally purified by MPLC to give amide or sulfonamide 9c. For examples of 9c wherein the side chain R1 or R9 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 9c was stirred in 30-80% TFA/CH2Cl2 for 1-3 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 9c. 
A solution of 1 mmol of N-Cbz-protected amino acid 10a and the appropriate amine (ZH, 1 mmol), diamine (ZH2, 0.5 mmol), or triamine (ZH3 0.33 mmol), was treated with 1.1 mmol of HOBt, 1.1 mmol of DIEA, and 2.1 mmol of EDCI in 3-6 mL of CH2Cl2 or DMF. [Alternatively, 1 mmol of the pentafluorophenyl ester or N-hydroxysuccinimide ester of 10a was mixed with the appropriate portion of amine (ZH), diamine (ZH2), or triamine (ZH3) in 3-6 mL of DMF.] The solution was stirred at room temperature under N2 for 12-24 h, and EtOAc was added. The organic solution was washed with 5% aqueous citric acid, water, saturated NaHCO3, and brine, dried over Na2SO4, and concentrated. The crude product was optionally purified by MPLC to afford amide 10b. Compound 10b was stirred in 30-80% TFA/CH2Cl2 for 1-3 h. The reaction mixture was concentrated to provide the TFA salt which was dissolved in CH2Cl2 or EtOAc and washed with aqueous NaOH or Na2CO3, dried over Na2SO4, and concentrated to give 10c as the free base.
A solution of 1 mmol of amino acid ester 10c (n=1), 2.5-3 mmol of the appropriately substituted chloride or bromide 2c, and 2.5-3 mmol of an appropriate base such as DIEA, Na2CO3, or Cs2CO3 in 3-5 mL of DMF was heated at 50-100xc2x0 C. under N2 for 18-24 h. (For examples of 10c where n=2 or 3, the amounts of 2c and base were increased by two- or three-fold, respectively.) The reaction mixture was cooled and partitioned between water and EtOAc. The organic layer was washed three times with water and once with brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure amide 10d.
Alternatively, a solution of 1 mmol of amino acid ester 10c (n=1), 2.5-3 mmol of the appropriately substituted aldehyde 2a, and 2.5-3 mmol of borane-pyridine complex in 3-5 mL of DMF or EtOH was stirred at room temperature under N2 for 3-5 days. (For examples of 10c where n=2 or 3, the amounts of 2c and borane-pyridine complex were increased by two- or three-fold, respectively.) The mixture was concentrated to dryness and was partitioned between water and CH2Cl2, washed with brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure amide 10d.
For examples of 10d where R1=CH2CH2CO2-t-Bu or CH2CO2-t-Bu, 10d was stirred in 30-80% TFA/CH2Cl2 for 1-24 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give acid 10e.
For examples of 10d where R1 is equal to (CH2)4NHBoc, 10d was stirred in 30-80% TFA/CH2Cl2 for 1-24 h. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give amine 10f as the TFA salt which was optionally dissolved in CH2Cl2 or EtOAc, washed with aqueous NaOH or Na2CO3, dried over Na2SO4, and concentrated to give 10f as the free base.
A solution of 1 mmol of 10f, 1-4 mmol of an appropriate base such as DIEA, and 1-2 mmol of the appropriately substituted cyclic anhydride 3e was stirred in CH2Cl2 or DMF under N2 overnight. The resulting mixture was diluted with CH2Cl2 or EtOAc and washed with aqueous HCl, water, and brine, was dried over Na2SO4, and concentrated to provide acid 10g. Alternatively, 1 mmol of 10f, 1-4 mmol of an appropriate base such as DIEA, and 1-2 mmol of the appropriately substituted carboxylic acid anhydride (R11CO)2O or acid chloride R11COCl was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 10h. Alternatively, 1 mmol of 8e, 14 mmol of an appropriate base such as DIEA, and 1-2 mmol of the appropriately substituted isocyanate R12NCO was stirred in CH2Cl2 or DMF under N2 overnight and worked up as above to provide 10i. 
An equivalent of an N-Fmoc-protected amino acid 11a which is bound to a polystyrene resin such as Wang resin is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. After filtration and washing with solvent, the resin is suspended in an appropriate solvent such as DMF. A solution of about 2-3 equivalents of an appropriately substituted halide 11b and a suitable base such DIEA is added to the resin bound amino acid and this mixture is shaken for 18-36 h. The resulting mixture is washed with several portions of a suitable solvent and is suspended and shaken in an acidic solution, such as 50% TFA/CH2Cl2, over several hours to cleave the acid from the resin to give a mixture of the Nxcex1,Nxcex1-bis-cinnamyl amino acid 11c and the Nxcex1-cinnamyl amino acid 11d.
By varying the resin bound amino acid 11a, one may obtain many of the compounds of the invention. The following resin bound amino acids may be used in Scheme 11: alanine, N-g-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)arginine, xcex2-(4-methyltrityl)asparagine, aspartic acid (xcex2-t-butyl ester), S-(trityl)cysteine, xcex3-(4-methyltrityl)glutamine, glutamic acid (xcex2-t-butyl ester), glycine, N-imidazolyl-(trityl)histidine, isoleucine, leucine, N-xcex5-(2-chlorobenzyloxycarbonyl)lysine, N-xcex5-(t-butoxycarbonyl)lysine, methionine, phenylalanine, proline, O-(t-butyl)serine, O-(t-butyl)threonine, N-indolyl-(t-butoxycarbonyl)tryptophan, O-(t-butyl)tyrosine, valine, xcex2-alanine, xcex1-aminoadipic acid, 4-aminobutanoic acid, 6-aminohexanoic acid, xcex1-aminosuberic acid, 5-aminopentanoic acid, p-aminophenylalanine, xcex1-aminopimelic acid xcex3-carboxyglutamic acid, p-carboxyphenylalanine, carnitine, citrulline, xcex1,xcex2-diaminopropionic acid, xcex1,xcex3-diaminobutyric acid, homocitrulline, homoserine, and statine. 
An equivalent of an N-Fmoc-protected amino acid which is bound to a resin 11a is suspended in a suitable solvent such as DMF. This solvent is removed and the nitrogen protecting group (Fmoc) is removed by stirring the resin bound amino acid with an organic base, such as piperidine, and an addition portion of the solvent. After filtration and washing with solvent, the resin is suspended in an appropriate solvent such as trimethyl orthoformate (TMOF), an appropriately substituted aldehyde 12a (5 equivalents) is added, and the mixture is shaken under N2 overnight. This mixture is treated with a suspension of NaBH(OAc)3 (5 equivalents) in CH2Cl2 and shaken under N2 overnight. After filtration and washing with a suitable solvent, the resulting product, resin bound Nxcex1-monosubstituted amino acid 12b, is suspended and shaken in an acidic solution, such as 50% TFA/CH2Cl2, over several hours to cleave the acid from the resin to give the Nxcex1-cinnamyl amino acid 11d.
The resin 12b is suspended in an appropriate solvent such as DMF and is filtered. The appropriately substituted halide 12c and an appropriate base such as DIEA are added with some additional solvent and the mixture is shaken under N2 for 18-36 h. The resin bound Nxcex1,Nxcex1-cinnamyl amino acid 12d is isolated from the suspension and the resin is cleaved with an acidic solution as described above to give the free acid 12e. 
A solution of of amino acid ester 13a, an appropriately substituted halide 11b, and an appropriate base such as DIEA, Na2CO3, or Cs2CO3 in a suitable solvent, such as DMF, is heated at 50-100xc2x0 C. under N2 overnight, or until the starting material is exhausted, to give a mixture of the Nxcex1,Nxcex1-bis-cinnamyl amino acid ester 13b and Nxcex1-cinnamyl amino acid ester 13c. If the side chain of R1 contains an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, those groups may be cleaved by treatment with an acidic solution such as 30-80% TFA/CH2Cl2 or 2-4N HCl in EtOAc. For examples of 13b and 13c where the ester group R4 is a primary alkyl group such as methyl or ethyl, esters 13b and 13c may be independently converted to the corresponding acids 11c and 11d by hydrolysis with an appropriate base such as aqueous NaOH, KOH, or LiOH. For examples of 13b and 13c where the ester group R4 is an acid-cleavable group such as t-butyl, esters 13b and 13c may be independently converted to the corresponding acids 11c and 11d by treatment with an acidic solution such as 30-80% TFA/CH2Cl2 or 2-4N HCl in EtOAc. 
A solution of 1 mmol of amino acid ester and 1-1.5 mmol of the appropriately substituted aldehyde 12a in 3-5 mL of TMOF was stirred at room temperature under N2 overnight. The solution was concentrated and used directly for the next reaction; optionally, the solution was partitioned between EtOAc and water, washed with brine, dried over Na2SO4, and concentrated to give crude product, which was purified by MPLC to give mono-substituted product 14a. For examples of 14a wherein the side chain R1 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 8c was treated with an acidic solution such as 30-80% TFA/CH2Cl2 or 2-4N HCl in EtOAc. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 14a. For examples of 14a where the ester group R4 is a primary alkyl group such as methyl or ethyl, esters 14a may be converted to the corresponding acids 11d by hydrolysis with an appropriate base such as aqueous NaOH, KOH, or LiOH. For examples of 14a where the ester group R4 is an acid-cleavable group such as t-butyl, esters 14a may be converted to the corresponding acids 11d by treatment with an acidic solution such as 30-80% TFA/CH2Cl2 or 2-4N HCl in EtOAc.
Amino ester 14a was dissolved in DMF, combined with 1.1-1.5 mmol of the appropriately substituted chloride or bromide 12c, and heated at 50-100xc2x0 C. overnight. The reaction mixture was cooled and partitioned between water and EtOAc. The organic layer was washed with water and brine, dried over Na2SO4, and concentrated. The crude product was purified by MPLC to give pure 14b. For examples of 14b wherein the side chain R1 contained an acid-cleavable protecting group such as t-butylcarbamate, t-butyl ester, or t-butyl ether, 8c was treated with an acidic solution such as 30-80% TFA/CH2Cl2 or 2-4N HCl in EtOAc. The reaction mixture was concentrated and optionally dissolved in HOAc and freeze-dried to give the deprotected form of 14b. For examples of 14b where the ester group R4 is a primary alkyl group such as methyl or ethyl, esters 14b may be converted to the corresponding acids 12e by hydrolysis with an appropriate base such as aqueous NaOH, KOH, or LiOH. For examples of 14b where the ester group R4 is an acid-cleavable group such as t-butyl, esters 14b may be converted to the corresponding acids 12e by treatment with an acidic solution such as 30-80% TFA/CH2Cl2 or 2-4N HCl in EtOAc. 
Although the claimed compounds are useful as competitive binders to the EPO receptor, some compounds are more active than others and are either preferred or particularly preferred. 
The particularly preferred xe2x80x9cR1xe2x80x9d s are the side chain of lysine, ornithine, arginine, aspartic acid, glutamic acid, glutamine, cysteine, methionine, serine, and threonine.
The particularly preferred xe2x80x9cR2 and R3xe2x80x9d s are phenoxy, substituted phenoxy, benzyloxy, and substituted benzyloxy.
The particularly preferred xe2x80x9cR4 and R5xe2x80x9d s are phenoxy, substituted phenoxy, benzyloxy, and substituted benzyloxy.
The particularly preferred xe2x80x9cWxe2x80x9d is xe2x80x94CHxe2x95x90CHxe2x80x94
The particularly preferred xe2x80x9cQxe2x80x9d is xe2x80x94CHxe2x95x90CHxe2x80x94
The particularly preferred xe2x80x9cXxe2x80x9d are C1-5alkenyl and CH2.
The particularly preferred xe2x80x9cYxe2x80x9d are C1-5alkenyl and CH2.
The particularly preferred xe2x80x9cnxe2x80x9d are 1 and 2.
The particularly preferred xe2x80x9cZxe2x80x9d are hydroxy, methoxy, phenethylamino, substituted phenethylamino, and xe2x80x94NH(CH2)2O(CH2)2O(CH2)2NHxe2x80x94.
Pharmaceutically useful compositions the compounds of the present invention, may be formulated according to known methods such as by the admixture of a pharmaceutically acceptable carrier. Examples of such carriers and methods of formulation may be found in Remington""s Pharmaceutical Sciences. To form a pharmaceutically acceptable composition suitable for effective administration, such compositions will contain an effective amount of the compound of the present invention.
Therapeutic or diagnostic compositions of the invention are administered to an individual in amounts sufficient to treat or diagnose disorders in which modulation of EPO receptor-related activity is indicated. The effective amount may vary according to a variety of factors such as the individual""s condition, weight, sex and age. Other factors include the mode of administration. The pharmaceutical compositions may be provided to the individual by a variety of routes such as subcutaneous, topical, transdermal, oral and parenteral.
The term xe2x80x9cchemical derivativexe2x80x9d describes a molecule that contains additional chemical moieties which are not normally a part of the base molecule. Such moieties may improve the solubility, half-life, absorption, etc. of the base molecule. Alternatively the moieties may attenuate undesirable side effects of the base molecule or decrease the toxicity of the base molecule. Examples of such moieties are described in a variety of texts, such as Remington""s Pharmaceutical Sciences.
Compounds disclosed herein may be used alone at appropriate dosages defined by routine testing in order to obtain optimal inhibition of the EPO receptor or its activity while minimizing any potential toxicity. In addition, co-administration or sequential administration of other agents may be desirable.
The present invention also has the objective of providing suitable topical, transdermal, oral, systemic and parenteral pharmaceutical formulations for use in the novel methods of treatment of the present invention. The compositions containing compounds according to this invention as the active ingredient for use in the modulation of EPO receptors can be administered in a wide variety of therapeutic dosage forms in conventional vehicles for administration. For example, the compounds or modulators can be administered in such oral dosage forms as tablets, capsules (each including timed release and sustained release formulations), pills, powders, granules, elixirs, tinctures, solutions, suspensions, syrups and emulsions, or by transdermal delivery or injection. Likewise, they may also be administered in intravenous (both bolus and infusion), intraperitoneal, subcutaneous, topical with or without occlusion, transdermal, or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. The compounds of the present invention may be delivered by a wide variety of mechanisms, including but not limited to, transdermal delivery, or injection by needle or needle-less injection means. An effective but non-toxic amount of the compound desired can be employed as an EPO receptor modulating agent.
The daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per patient, per day. For oral administration, the compositions are preferably provided in the form of scored or unscored tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, and 50.0 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. An effective amount of the drug is ordinarily supplied at a dosage level of from about 0.0001 mg/kg to about 100 mg/kg of body weight per day. The range is more particularly from about 0.001 mg/kg to 10 mg/kg of body weight per day. The dosages of the EPO receptor modulators are adjusted when combined to achieve desired effects. On the other hand, dosages of these various agents may be independently optimized and combined to achieve a synergistic result wherein the pathology is reduced more than it would be if either agent were used alone.
Advantageously, compounds or modulators of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, compounds or modulators for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
For combination treatment with more than one active agent, where the active agents are in separate dosage formulations, the active agents can be administered concurrently, or they each can be administered at separately staggered times.
The dosage regimen utilizing the compounds or modulators of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound thereof employed. A physician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition. Optimal precision in achieving concentrations of drug within the range that yields efficacy without toxicity requires a regimen based on the kinetics of the drug""s availability to target sites. This involves a consideration of the distribution, equilibrium, and elimination of a drug.
In the methods of the present invention, the compounds or modulators herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as xe2x80x9ccarrierxe2x80x9d materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include, without limitation, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
For liquid forms the active drug component can be combined in suitably flavored suspending or dispersing agents such as the synthetic and natural gums, for example, tragacanth, acacia, methyl-cellulose and the like. Other dispersing agents which may be employed include glycerin and the like. For parenteral administration, sterile suspensions and solutions are desired. Isotonic preparations which generally contain suitable preservatives are employed when intravenous administration is desired.
Topical preparations containing the active drug component can be admixed with a variety of carrier materials well known in the art, such as, e.g., alcohols, aloe vera gel, allantoin, glycerine, vitamin A and E oils, mineral oil, PPG2 myristyl propionate, and the like, to form, e.g., alcoholic solutions, topical cleansers, cleansing creams, skin gels, skin lotions, and shampoos in cream or gel formulations.
The compounds or modulators of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds or modulators of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinyl-pyrrolidone, pyran copolymer, polyhydroxypropylmethacryl-amidephenol, polyhydroxy-ethylaspartamidephenol, or polyethyl-eneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds or modulators of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydro-pyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels, and other suitable polymers known to those skilled in the art.
For oral administration, the compounds or modulators may be administered in capsule, tablet, or bolus form or alternatively they can be mixed in the animals feed. The capsules, tablets, and boluses are comprised of the active ingredient in combination with an appropriate carrier vehicle such as starch, talc, magnesium stearate, or di-calcium phosphate. These unit dosage forms are prepared by intimately mixing the active ingredient with suitable finely-powdered inert ingredients including diluents, fillers, disintegrating agents, and/or binders such that a uniform mixture is obtained. An inert ingredient is one that will not react with the compounds or modulators and which is non-toxic to the animal being treated. Suitable inert ingredients include starch, lactose, talc, magnesium stearate, vegetable gums and oils, and the like. These formulations may contain a widely variable amount of the active and inactive ingredients depending on numerous factors such as the size and type of the animal species to be treated and the type and severity of the infection. The active ingredient may also be administered as an additive to the feed by simply mixing the compound with the feedstuff or by applying the compound to the surface of the feed. Alternatively the active ingredient may be mixed with an inert carrier and the resulting composition may then either be mixed with the feed or fed directly to the animal. Suitable inert carriers include corn meal, citrus meal, fermentation residues, soya grits, dried grains and the like. The active ingredients are intimately mixed with these inert carriers by grinding, stirring, milling, or tumbling such that the final composition contains from 0.001 to 5% by weight of the active ingredient.
The compounds or modulators may alternatively be administered parenterally via injection of a formulation consisting of the active ingredient dissolved in an inert liquid carrier. Injection may be either intramuscular, intraruminal, intratracheal, or subcutaneous, either by needle or needle-less means. The injectable formulation consists of the active ingredient mixed with an appropriate inert liquid carrier. Acceptable liquid carriers include the vegetable oils such as peanut oil, cotton seed oil, sesame oil and the like as well as organic solvents such as solketal, glycerol formal and the like. As an alternative, aqueous parenteral formulations may also be used. The vegetable oils are the preferred liquid carriers. The formulations are prepared by dissolving or suspending the active ingredient in the liquid carrier such that the final formulation contains from 0.005 to 10% by weight of the active ingredient.
Topical application of the compounds or modulators is possible through the use of a liquid drench or a shampoo containing the instant compounds or modulators as an aqueous solution or suspension. These formulations generally contain a suspending agent such as bentonite and normally will also contain an antifoaming agent. Formulations containing from 0.005 to 10% by weight of the active ingredient are acceptable. Preferred formulations are those containing from 0.01 to 5% by weight of the instant compounds or modulators.
The compounds of Formula I may be used in pharmaceutical compositions to treat patients (humans and other mammals) with disorders or conditions associated with the production of erythropoietin or modulated by the EPO receptor. The compounds can be administered in the manner of the commercially available product or by any oral or parenteral route (including but not limited to, intravenous, intraperitoneal, intramuscular, subcutaneous, dermal patch), where the preferred route is by injection. When the method of administration is intravenous infusion, compound of Formula I may be administered in a dose range of about 0.01 to 1 mg/kg/min. For oral administration, the dose range is about 0.1 to 100 mg/kg.
The pharmaceutical compositions can be prepared using conventional pharmaceutical excipients and compounding techniques. Oral dosage forms may be used and are elixirs, syrups, capsules, tablets and the like. Where the typical solid carrier is an inert substance such as lactose, starch, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, mannitol and the like; and typical liquid oral excipients include ethanol, glycerol, water and the like. All excipients may be mixed as needed with disintegrants, diluents, granulating agents, lubricants, binders and the like using conventional techniques known to those skilled in the art of preparing dosage forms. Parenteral dosage forms may be prepared using water or another sterile carrier.
Typically the compounds of Formula I are isolated as the free base, however when possible pharmaceutically acceptable salts can be prepared. Examples of such salts include hydrobromic, hydroiodic, hydrochloric, perchloric, sulfuric, maleic, fumaric, malic, tartaric, citric, benzoic, mandelic, methanesulfonic, hydroethanesulfonic, benzenesulfonic, oxalic, pamoic, 2-naphthalenesulfonic, p-toluenesulfonic, cyclohexanesulfamic and saccharic.
In order to illustrate the invention the following examples are included. These examples do not limit the invention. They are only meant to suggest a method of practicing the invention. Those knowledgeable in chemical synthesis and the treatment of EPO related disorders may find other methods of practicing the invention. However those methods are deemed to be within the scope of this invention.